Fig. 8. Effects of Gd³⁺ on vinculin and phosphotyrosine organization. NIH3T3 cells were treated with 100 µM Gd³⁺ (A-E) or control buffer (F-J) for 30 minutes before processing for immunofluorescent staining of vinculin (A,F), actin filaments (B,G), or phosphotyrosine (E,J; in different cells). C and H show combined actin-vinculin images. The control cell showed the characteristic localization of vinculin at focal adhesions (F), which were concentrated at the termini of stress fibers (H) and appeared as dark plaques in the IRM image (I). The Gd³⁺-treated cell showed a reduction in stress fiber number and intensity, and an increase in cortical actin intensity (B). Short stress fibers persist near the nucleus. Vinculin localization was much reduced at focal adhesions, and appeared primarily perinuclear (A). Staining for phosphotyrosine showed a similar decrease at focal adhesions in Gd³⁺ treated cells (E). IRM image of treated cell showed adhesion plaques throughout the cell (D), which appeared less elongated than those in control cells (I). Scale bar: 20 µm.